Abstract
The Drosophila genes Pig-1 and Sgs-4 are adjacent and oppositely transcribed genes which share a common enhancer region located in the intergenic region between the two transcription start sites. Both genes are transcribed in the same tissue, the larval salivary gland, but they are expressed at different times, the expression switching from Pig-1 to Sgs-4 expression during development of the third-instar larva. Here we describe partial purification and functional analysis of a DNA-binding protein, secretion enhancer binding protein 1 (SEBP1), from nuclear extracts of whole Drosophila larvae. SEBP1 binds to a single site within the Pig-1/Sgs-4 intergenic region. Point mutations that disrupt this site had no significant effect on activation of the Sgs-4 promoter in constructs lacking the Pig-1 promoter. However, if the Pig-1 promoter was present, the same mutations led to a strong attenuation of Sgs-4 promoter activity, suggesting that binding of SEBP1 is necessary for the Pig-1/Sgs-4 transcriptional switch. The data are consistent with a model in which the SEBP1 binding site acts as a transient silencing element that facilitates the reorganization of promoter-enhancer interactions by disrupting pre-existing interactions, thus enabling stage-specific activators of Sgs-4 to bind to their enhancer target sites.
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